PROJECT SUMMARY ? FLOW CYTOMETRY SHARED RESOURCE The Flow Cytometry Shared Resource?s (FCSR) primary goal is to provide support for the use of flow cytometric services by Cancer Center investigators and, regionally, to the greater basic research community. The specific aims are to: (i) provide state-of-the-art technological capability for high-quality, multi-parameter analyses and/or sorting of different cell types from homogeneous or mixed cell populations via staff or user- friendly operation; (ii) provide training and ongoing support in basic and advanced user-friendly operations, as well as expertise to assist investigators in choosing reagents or instrument conditions that optimize the use of cytometry for their experimental needs; (iii) advise and provide technical support for analysis of flow cytometry data for publication, research presentations, and grant applications, along with management of cytometric data. The services provided by the facility include sample acquisition and analysis, cell sorting, on-site instrument maintenance, training, advice, and consultation. Currently, the FCSR houses a Becton-Dickinson (BD) FACSAria II high-speed cell sorter (installed 2009), a MoFlo Astrios EQ high-speed cell sorter (2014), and the following user-friendly analysis instruments: 2 BD LSRIIs (18-color and 14-color), a BD FACSCelesta with a High Throughput Sampler (HTS) (12-color, 2016), and a BD FACSCalibur with a HTS. In addition, the FCSR provides access to an Amnis ImageStreamx imaging cytometry system. The FCSR physical plant includes additional workstations, a library of flow cytometry journals, protocol guides, and other references. Twenty-four- hour access is available for all investigator-operated analysis instruments, reserved through an on-line calendar system. In the last Cancer Center Support Grant budget cycle period, there were substantial improvements to equipment and infrastructure: first, the FCSR was relocated to a single laboratory of 1,137 NSF located in the west wing of the Cancer Center with ease of access for all Cancer Center investigators; second, the FCSR acquired a new cell sorter, a 7-laser, 32-parameter MoFlo AstriosEQ (NIH shared instrumentation grant, 1S10 OD017998, PI, Dr. Montaner), and a 3-laser, 12-color FACSCelesta analysis instrument (Institute-funded). During the past project period, Cancer Center members from all Programs have extensively utilized the FCSR to accomplish pivotal research projects and increase the priority and impact of grant applications and peer-reviewed publications. Ongoing improvements in the FCSR enable Cancer Center members to accomplish rapid, highly automated analyses of diverse cell populations for better phenotypical characterization of cellular subsets and their functional characterization. This is especially valuable in analyzing populations of blood or immune-system cells, as well as expression of fluorescent protein-expressing vectors. Cell sorting permits the sterile purification of these cells for subsequent experimentation.